Articles
Optimizing regeneration systems for highbush blueberries
Article number
1440_51
Pages
369 – 376
Language
English
Abstract
Tissue culture and regeneration limitations pose substantial challenges to advancing (Vaccinium spp.) cultivar development, restricting the widespread use of tissue culture techniques in commercial production.
This highlights the critical need for an optimized and standardized regeneration protocol.
To address this, we established improved in vitro regeneration protocols for six highbush blueberry (Vaccinium corymbosum L.) cultivars: ‘Emerald’, ‘Rowan’ (‘NC 3104’), ‘NC 5288’, ‘Legacy’, ‘Pinnacle’, and ‘Jewel’, which exhibit varying degrees of tissue culture recalcitrance.
We assessed the impact of the auxin 1-naphthaleneacetic acid (NAA) and cytokinins, including zeatin (ZT), thidiazuron (TDZ), 1-(2-Chloro-4-pyridyl)-3-phenylurea (CPPU), and 6-(gamma, gamma-Dimethylallylamino) purine (2iP), on callus formation and regeneration from leaf explants.
Media supplemented with zeatin and 2iP yielded the highest regeneration rates across cultivars, reaching 53.33, 61.67, 60, 65, and 43.33%, respectively.
This cytokinin combination also promoted superior callus induction, higher shoot proliferation, and more efficient callus-to-shoot conversion, demonstrating its broad effectiveness as a regeneration medium.
These findings contribute valuable refinements to tissue culture methodologies, laying a strong foundation for the development of enhanced blueberry cultivars and the application of genome editing technologies.
Notably, this study is the first to report in vitro regeneration protocols for ‘Rowan’ (‘NC3104’), ‘NC 5288’, and ‘Pinnacle’, expanding opportunities for blueberry genetic improvement.
This highlights the critical need for an optimized and standardized regeneration protocol.
To address this, we established improved in vitro regeneration protocols for six highbush blueberry (Vaccinium corymbosum L.) cultivars: ‘Emerald’, ‘Rowan’ (‘NC 3104’), ‘NC 5288’, ‘Legacy’, ‘Pinnacle’, and ‘Jewel’, which exhibit varying degrees of tissue culture recalcitrance.
We assessed the impact of the auxin 1-naphthaleneacetic acid (NAA) and cytokinins, including zeatin (ZT), thidiazuron (TDZ), 1-(2-Chloro-4-pyridyl)-3-phenylurea (CPPU), and 6-(gamma, gamma-Dimethylallylamino) purine (2iP), on callus formation and regeneration from leaf explants.
Media supplemented with zeatin and 2iP yielded the highest regeneration rates across cultivars, reaching 53.33, 61.67, 60, 65, and 43.33%, respectively.
This cytokinin combination also promoted superior callus induction, higher shoot proliferation, and more efficient callus-to-shoot conversion, demonstrating its broad effectiveness as a regeneration medium.
These findings contribute valuable refinements to tissue culture methodologies, laying a strong foundation for the development of enhanced blueberry cultivars and the application of genome editing technologies.
Notably, this study is the first to report in vitro regeneration protocols for ‘Rowan’ (‘NC3104’), ‘NC 5288’, and ‘Pinnacle’, expanding opportunities for blueberry genetic improvement.
Publication
Authors
F. Shepard, W. Liu, R. Dewey, K. Da, D. Reiland, C. Almeyda-Becerra, H. Ashrafi
Keywords
blueberry, Vaccinium corymbosum, genotype-independent regeneration, tissue culture recalcitrance, 2iP, zeatin
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